Fig. 1: CLOCK and BMAL1 enhance the formation of F-actin.
From: CLOCK and BMAL1 stabilize and activate RHOA to promote F-actin formation in cancer cells
a, b F-actin was stained with TRITC-phalloidin (Red) in HepG2 cells transfected with HA-tagged BMAL1 (BMAL1+), HA-tagged CLOCK (CLOCK+), shRNA for BMAL1 (shBMAL1), and CLOCK (shCLOCK) vectors. Nuclei were stained with DAPI. The scale bars represent 10 μm. The F-actin levels were quantified with the fluorescence intensity ratio using ImageJ. The fluorescence intensity of the control was taken as 1. Data were analyzed using a t test (n = 3) and presented as the mean ± sd; *p < 0.05; **p < 0.01. c F-actin and G-actin from control, shBMAL1, BMAL1+, shCLOCK, and CLOCK+ HeLa cells were separated by ultraspeed centrifugation and analyzed by western blotting using an antibody against β-actin. The mean ratio of F-actin to G-actin and SD were plotted, and a t test was performed (n = 3); n.s. not significant; *p < 0.05; **p < 0.01. d, e The expression levels of CLOCK and BMAL1 were verified by western blotting in HeLa and HepG2 cells transfected with the corresponding plasmids, respectively. GAPDH was used as a loading control, and the protein level of the control was taken as 1. Data were analyzed using a t test (n = 3) and presented as the mean ± sd; *p < 0.05; **p < 0.01
